Localization of host immunoglobulin G to the surface of Pneumocystis carinii.
In immunoblotting studies of Pneumocystis carinii surface proteins, we found that a secondary antibody, anti-human immunoglobulin G (IgG), recognized a 52-kilodalton (kDa) band in homogenates of P. carinii purified from human autopsy lungs and bronchoalveolar lavage fluids, even when serum as a source of primary antibody was omitted. The electrophoretic mobility of the 52-kDa band is identical to that of IgG heavy chains. In addition to affinity-purified, anti-human IgG, monoclonal antibodies specific for the Fab and Fc regions of human IgG recognized the 52-kDa band. To determine whether the 52-kDa band represents IgG bound to the surface of P. carinii, we treated intact organisms with Triton X-100 and acid in order to elute immunoglobulin from the surface of P. carinii. After purification over a protein G column, the eluate comigrated with human IgG, was recognized by anti-IgG, and bound to discrete bands with molecular sizes of 65 to 70, 60, 50, and 35 kDa in purified, rat-derived P. carinii. To confirm the presence of human IgG on the surface of P. carinii, we performed immunocytochemical and immunoelectron microscopic studies. Staining of intact P. carinii aggregates by anti-human IgG was pronounced and was abolished by acid treatment. IgA was also present. Ultrastructural studies showed the presence of IgG on the cyst wall and on fine membranous structures and vesicles adjacent to cysts. We conclude that the surface of P. carinii is coated with human IgG. The close association of human IgG with P. carinii may have implications for the pathogenesis of P. carinii pneumonia in acquired immunodeficiency syndrome.
Blumenfeld
W
W
Center for Immunochemistry, Veterans Administration Medical Center, San Francisco, California 94121.
Mandrell
R E
RE
Jarvis
G A
GA
Griffiss
J M
JM
D000163:Acquired Immunodeficiency Syndrome / Q000150:complications; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000032:analysis* / Q000276:immunology; D007150:Immunohistochemistry; D011010:Pneumocystis / Q000276:immunology*; D011020:Pneumonia, Pneumocystis / Q000209:etiology; D011947:Receptors, Antigen, B-Cell / Q000032:analysis*
D007074:Immunoglobulin G; D011947:Receptors, Antigen, B-Cell